Method for analyzing color code encoded in magnetic structure

ABSTRACT

Provided is a color encoding method including providing a composition including a liquid medium and magnetic nanoparticles dispersed in the liquid medium; applying a magnetic field to the composition to align the magnetic nanoparticles; and applying a patterned energy source to the composition to solidify the composition, wherein more than one region of the composition are sequentially solidified with varying magnetic field strength to fix a plurality of color codes.

CROSS REFERENCE TO PRIOR APPLICATIONS

This application claims the benefit under 35 U.S.C. §120 and §365(c) toa prior PCT International Patent Application No. PCT/KR2010/002249(filed on Apr. 13, 2010 and designating the U.S.), which claims priorityto U.S. Provisional Application No. 61/169,260 (filed on Apr. 14, 2009)and Korean Patent Application No. 10-2010-0029613 (filed on Mar. 31,2010), which are all hereby incorporated by reference in their entirety.

TECHNICAL FIELD

The described technology generally relates to a color encoded magneticstructure.

BACKGROUND

Multiplex analysis based on encoded particles has attracted greatattention in high throughput biomolecule screening such as drugdiscovery and clinical diagnostics due to its expandability and fastreactivity. The multiplex analysis may be accomplished by mixing manyencoded probe particles together in the same space containing targetanalytes.

In various samples, it is necessary to use a large number of distinctcodes to increase throughput. To ensure a sufficiently large number ofcodes for microparticles, a spectral encoding method using coloringmaterials such as quantum dots or fluorescent dyes, which are embeddedinside or attached on the surface of the microparticle and a graphicalencoding method based on patterning of an optically detectable elementson the surface of the microparticle have been suggested.

SUMMARY

In one embodiment, a color encoding method is provided. The methodincludes: providing a composition including a liquid medium and magneticnanoparticles dispersed in the liquid medium; applying a magnetic fieldto the composition to align the magnetic nanoparticles; and applying apatterned energy source to the composition to solidify the composition,wherein more than one region of the composition are sequentiallysolidified with varying magnetic field strength to fix a plurality ofcolor codes.

In another embodiment, a method of manufacturing a color encodedmagnetic structure is provided. The method includes: filling amicrofluidic channel with a composition including a curable material andmagnetic nanoparticles dispersed in the curable material; applying amagnetic field to the composition in the microfluidic channel to formone-dimensional (1D) chain structures of the magnetic nanoparticles; andapplying patterned UV rays to the composition to form a free-floatingparticle with fixed 1D chain structures.

In yet another embodiment, a color encoded magnetic structure isprovided. The color encoded magnetic structure includes: a solid medium;and a code region present in the solid medium and including magneticnanoparticles aligned in a chain structure. The color encoded magneticstructure is encoded in multilevel by generating structural colorsdetermined by inter-particle distance between the aligned magneticnanoparticles.

In still another embodiment, a method of analyzing a color code isprovided. The method includes: providing a magnetic structure includinga probe region and a code region encoded with a color code; binding atarget with the probe region; and decoding information from the magneticstructure with the target, wherein the code region includes a photoniccrystal of magnetic nanoparticles.

The Summary is provided to introduce a selection of concepts in asimplified form that are further described below in the DetailedDescription. The Summary is not intended to identify key features oressential features of the claimed subject matter, nor is it intended tobe used as an aid in determining the scope of the claimed subjectmatter.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

The above and other objects, features and advantages of the presentdisclosure will become more apparent to those of ordinary skill in theart by describing in detail exemplary embodiments thereof with referenceto the attached drawings in which:

FIG. 1 is a diagram showing an exemplary embodiment of a composition forcolor encoding;

FIG. 2 is a diagram for explaining a principle of generating astructural color using a composition for color encoding;

FIG. 3 is a schematic diagram showing a procedure of generatingmulticolor encoded microparticles;

FIG. 4 is a schematic diagram viewed from a cross-sectional view takenalong line A-A′ of FIG. 3;

FIG. 5 shows images of various types of color encoded microparticles;

FIG. 6 shows images comparing a conventional binary encoded particlewith a color encoded particle;

FIG. 7 is a microscope image of a color encoded magnetic structure;

FIG. 8 shows reflective microscope images of multiplexed assays based onhybridization of three kinds of encoded particles and DNA oligomertargets;

FIG. 9 shows results obtained after an encoded particle having a probe 1is hybridized with a complementary oligomer target (T1); and

FIG. 10 shows results obtained after the encoded particle of FIG. 9 isdecoded.

DETAILED DESCRIPTION

It will be readily understood that the components of the presentdisclosure, as generally described and illustrated in the Figuresherein, could be arranged and designed in a wide variety of differentconfigurations. Thus, the following more detailed description of theembodiments of apparatus and methods in accordance with the presentdisclosure, as represented in the Figures, is not intended to limit thescope of the disclosure, as claimed, but is merely representative ofcertain examples of embodiments in accordance with the disclosure. Thepresently described embodiments will be best understood by reference tothe drawings, wherein like parts are designated by like numeralsthroughout. Moreover, the drawings are not necessarily to scale, and thesize and relative sizes of the layers and regions may have beenexaggerated for clarity.

As used in the description herein and throughout the claims, thefollowing terms take the meanings explicitly associated herein, unlessthe context clearly dictates otherwise: the meaning of “a”, “an”, and“the” includes plural reference, the meaning of “in” includes “in” and“on”. It will also be understood that when an element or layer isreferred to as being “on” another element or layer, the element or layermay be directly on the other element or layer or intervening elements orlayers may be present. As used herein, the term “and/or” may include anyand all combinations of one or more of the associated listed items.

According to one exemplary embodiment, color encoding may be performedby the following method: First, a composition including a liquid mediumand magnetic nanoparticles dispersed in the liquid medium is prepared.The liquid medium may be a photocurable material. In addition, themagnetic nanoparticles may include superparamagnetic materials. Themagnetic nanoparticle may be coated with a shell layer to improvedispersibility and solvation repulsion.

Subsequently, the magnetic nanoparticles are aligned by applying amagnetic field to the composition. Concurrently, the composition issolidified by irradiating a patterned energy source. Here, a colorencoded structure may be prepared by fixing a plurality of color codesby sequentially solidifying different regions of the composition withvarying the magnetic field strength. The patterned energy sourcesuitable for the solidification may include heat, UV rays, visible rays,infrared rays, an electron beam, etc. without limitation. The 1D chainstructure may be made by the alignment of the magnetic nanoparticles,and structural colors may be determined by inter-particle distancebetween the magnetic nanoparticles forming the chain structures. With anincrease in the magnetic field strength, a structural color of a shorterwavelength region may be produced. The composition may further include ahydrogen bonding solvent to form a solvation layer around the magneticnanoparticles.

For example, the irradiation of the patterned UV rays may be performedusing a physical mask or digital micromirror device (DMD). As apatterning method, a technique such as optofluidal maskless lithography(OFML) may be used to prevent diffusion of free radicals and produce ahigh-definition microscale pattern during polymerization.

The plurality of color codes may include information such as a shape, aposition and a color independently or in combination. For example, theplurality of color codes may be discriminated each other by theirspecific colors at different positions. The plurality of color codes maybe in the form of dots, lines or any other shapes. Since a photoniccrystal structure formed by the magnetic nanoparticles is maintained bythe solidification, a structural color from each color code may not bechanged and thus may be fixed.

FIG. 1 is a diagram showing an exemplary embodiment of a composition forcolor encoding. Referring to FIG. 1, the composition 100 for colorencoding may include a curable material 110 and magnetic nanoparticles120 dispersed in the curable material 110.

The magnetic nanoparticles 120 may include a cluster 122 of magneticnanocrystals. The size of the magnetic nanoparticles 120 may be severaltens to hundreds of nanometers, and the size of the magneticnanocrystals may be several to several tens of nanometers. Examples ofthe magnetic nanocrystals may include a magnetic materials or a magneticalloys. The magnetic material or magnetic alloy may include at least oneselected from the group consisting of Co, Fe₂O₃, Fe₃O₄, CoFe₂O₄, MnO,MnFe₂O₄, CoCu, CoPt, FePt, CoSm, NiFe and NiFeCo.

The magnetic nanoparticles 120 may be superparamagnetic nanoparticlesincluding a superparamagnetic material. The superparamagnetic materialhas magnetism only in the presence of an external magnetic field, unlikea ferromagnetic material in which magnetism can be maintained without amagnetic field. Usually, when the particle size of a ferromagneticmaterial is several to several hundreds of nanometers, the ferromagneticmaterial may be phase-changed into a superparamagnetic material. Forexample, when iron oxide is in the size of approximately 10 nm, it mayhave superparamagnetism.

In addition, the magnetic nanoparticles 120 may be, as shown in FIG. 1,coated with a shell layer 124 surrounding a core formed in the cluster122 of magnetic nanocrystals. The shell layer 124 allows the magneticnanoparticles 120 to be evenly distributed in the curable material 110.Furthermore, to be described later, the shell layer 124 may stimulatesolvation repulsion on a surface of each magnetic nanoparticle 120 tooffset potent magnetic attraction between the magnetic nanoparticles120. For example, the shell layer 124 may include silica. When the shelllayer 124 is surface-modified with silica, a known sol-gel process maybe used.

In addition, the composition 100 for color encoding may further includea hydrogen bonding solvent. As the hydrogen bonding solvent, variousalkanol solvents such as ethanol, isopropyl alcohol and ethylene glycolmay be used. Also, a solvation layer 126 surrounding the magneticnanoparticle 120 may be formed. For example, as the solvation layer 126is formed due to an influence of a silanol (Si—OH) functional group on asurface of the shell layer 124 having silica, a repulsion force betweenthe magnetic nanoparticles 120 may be induced. According to oneexemplary embodiment, the shell layer 124 and/or the solvation layer 126may not be present on the magnetic nanoparticles 120. In this case, anelectrostatic force on the surface of the magnetic nanoparticles 120 mayact as a repulsion force.

As the magnetic nanoparticles 120 are mixed with the curable material110 and subjected to mechanical stirring or ultrasonic treatment, thecomposition for color encoding 100 may be prepared. The magneticnanoparticles 120 may be included in the curable material 110 at avolume fraction of, for example, 0.01% to 20%. When the volume fractionof the magnetic nanoparticles 120 is less than 0.01%, reflectivity maybe decreased, and when the volume fraction of the magnetic nanoparticles120 is more than 20%, reflectivity may not be increased any more.

The curable material 110 may serve as a dispersion medium stablydispersing the magnetic nanoparticles 120 forming a photonic crystal. Inaddition, as the inter-particle distance between the magneticnanoparticles 120 is fixed by crosslinking of the curable material 110,a certain structural color may be continuously maintained after amagnetic field is eliminated.

The curable material 110 may include a liquid-phase material such as amonomer, an oligomer or a polymer having a crosslinkable site for curingreaction. The curable material 110 may include a liquid-phasehydrophilic polymer capable of forming a hydrogel. A hydrophilic polymeris a polymer suitable for dispersing the magnetic nanoparticles 120 dueto its hydrophilic groups. When the hydrophilic polymer is crosslinkedby an appropriate energy source, thereby forming a hydrogel having athree-dimensional network structure, the magnetic nanoparticles 120 maybe fixed.

Examples of the curable material 110 capable of forming a hydrogel mayinclude a silicon-containing polymer, polyacrylamide, polyethyleneoxide, polyethylene glycol diacrylate, polypropylene glycol diacrylate,polyvinylpyrrolidone, polyvinyl alcohol, polyacrylate or a copolymerthereof. For example, since the curable material 110, polyethyleneglycol diacrylate (PEG-DA), has an acrylate functional group at bothterminal ends of polyethylene glycol (PEG), the curable material 110 maybe crosslinked into a three-dimensional hydrogel via free radicalpolymerization. The curable material 110 may further include any type ofmedium which can be changed into a solid from a liquid.

The curable material 110 may further include an initiator, and theinitiator may induce free radical polymerization by an external energysource. The initiator may be an azo-based compound or a peroxide. Thecurable material 110 may further include a proper crosslinking agent,for example, N,N′-methylenebisacrylamide, methylenebismethacrylamide,ethylene glycol dimethacrylate, etc. The magnetic nanoparticles 120 maybe aligned in the curable material 110 to generate structural colorsunder an external magnetic field.

FIG. 2 is a diagram for explaining a principle of generating astructural color using a composition for color encoding. Referring toFIG. 2, when a magnetic field is not applied, the magnetic nanoparticles120 are randomly dispersed in the curable material 110, but when amagnetic field is applied from a nearby magnet, the magneticnanoparticles 120 may be aligned parallel to a direction of the magneticfield to form a photonic crystal, thereby emitting a structural color.The magnetic nanoparticles 120 aligned by the magnetic field may returnto the non-aligned state when the magnetic field is eliminated. Aphotonic crystal is a material having a crystal structure capable ofcontrolling light. Photons (behaving as waves) propagate through thisstructure—or not—depending on their wavelength. Wavelengths of lightthat are allowed to travel are known as modes, and groups of allowedmodes form bands. Disallowed bands of wavelengths are called photonicband gaps. This gives rise to distinct optical phenomena such asinhibition of spontaneous emission, high-reflecting omni-directionalmirrors and low-loss-waveguiding, amongst others. The magneticnanoparticles 120 present in a colloidal state may have an attractiveinteraction therebetween in the curable material 110 due to themagnetism when a magnetic field is applied outside, and also have arepulsive interaction caused by an electrostatic force and a solvationforce. By the balance between the attraction and the repulsion, themagnetic nanoparticles 120 are aligned at regular intervals, therebyforming a chain structure. Therefore, a inter-particle distance dbetween the aligned magnetic nanoparticles 120 may be determined by themagnetic field strength. As the magnetic field is stronger, theinter-particle distance d between the magnetic nanoparticles 120 alignedalong the direction of the magnetic field may be reduced. Theinter-particle distance d may be several to several hundreds ofnanometers with the magnetic field strength. With a lattice spacing inthe photonic crystal is changed, the wavelength of reflected light maybe changed according to Bragg's law. As the magnetic field strength isincreased, a structural color of a shorter wavelength region may begenerated. As a result, a wavelength of the reflected light may bedetermined by the strength of a specific magnetic field. Unlike theconventional photonic crystal reflected only at a certain wavelength,the photonic crystal may exhibit an optical response that is fast,extensive and reversible with respect to an external magnetic field. Asthe lattice spacing is changed with the variation in the nearby magneticfield, the reflective light with a specific wavelength may be inducedfrom external incident light.

According to an exemplary embodiment, a method of manufacturing a colorencoded magnetic structure using OFML is provided. The method mayinclude: filling a microfluidic channel with a composition including acurable material and magnetic nanoparticles dispersed in the curablematerial; applying a magnetic field to the composition in themicrofluidic channel to form one-dimensional (1D) chain structures ofthe magnetic nanoparticles; and applying patterned UV rays to thecomposition to form a free-floating particle with fixed 1D chainstructures.

The curable material may include a liquid-phase material such as amonomer, an oligomer or a polymer having a crosslinkable site. Thefree-floating particle may be encoded with a structural color generatedfrom the 1D chain structures.

The method of manufacturing a color encoded magnetic structure mayfurther include changing the magnetic field strength and sequentiallysolidifying more than one region of the composition to implementmultilevel coding. The composition flowing through the microfluidicchannel may be encoded with multicolored patterns by changing maskpatterns and sequentially irradiating the patterned UV rays to thecomposition.

The free-floating particles showing structural colors may include aplurality of color codes, respectively. The plurality of color codes mayhave various colors determined by the magnetic field strength at thetime of curing. A shape and colored pattern of the free-floatingparticle may be fixed by irradiation of the patterned UV rays.

FIG. 3 is a schematic diagram showing a procedure of generatingmulticolor encoded microparticles. In addition, FIG. 4 is a schematicdiagram viewed from a cross-sectional view taken along line A-A′ of FIG.3. Referring to FIGS. 3 and 4, a sequential process includingcooperation of magnetic field modulation and spatially controlled UVexposure is used. First, a polydimethylsiloxane (PDMS) micro fluidicchannel is filled with a composition including a photocurable resin andsuperparamagnetic nanoparticles dispersed in the photocurable resin.

Subsequently, a color of the composition is tuned by modulating anexternal magnetic field. A periodicity of the 1D chain structure of thesuperparamagnetic nanoparticles is changed with the magnetic fieldstrength, and light of corresponding wavelength is reflected. In ascanning electron microscope (SEM) image of the second step of FIG. 4,the superparamagnetic nanoparticles are aligned in a chain structure toform a completely reversible 1D photonic crystal (scale bar: 1 μm). Astronger magnetic field results in a shorter inter-particle distance,which corresponds to a shorter diffracted wavelength. Once a structuralcolor is determined by the external magnetic field, locally-patterned UVrays are irradiated to a specific region of the composition to fix thestructural color. A DMD may be used as a dynamic mask to irradiate thepatterned UV rays. The DMD may serve as a computer-controlled spatiallight modulator. In FIG. 3, the UV rays are reflected from the DMD andtherefore patterned UV rays are created. After the irradiation of thepatterned LTV rays, the specific region is encoded with a specific colorto generate an encoded bit.

Next encoded bits may be continuously created by sequentially changingthe magnetic field strength and the pattern of the DMD. Since colortuning and fixing process for each bit takes approximately several tensof a second, overall process can be swiftly performed. In addition,realignment procedure required for the process using general masks isnot needed, and thus the process is simple and less expensive. Moreover,an oxygen lubricant layer in the PDMS channel allows microparticlescreated by radical photopolymerization to move along the flow of a fluidwithout attachment to the PDMS channel wall.

By using such a characteristic, encoded particles having various colorsand shapes may be created under various levels of the magnetic field,together with the patterned UV, using OFML. That is, free-floatingparticles may be formed by injecting a liquid phase curable resincontaining a photonic crystal of magnetic nanoparticles into amicrofluidic channel, and performing in-situ photopolymerization inducedby patterned UV rays under various magnetic fields. The free-floatingparticles may be designed in desired shapes. For example, thefree-floating particles of rod-type or disc-type (e.g. circle, square,hexagon, octagon and so on) may be produced under various UV patterns.Different kinds of encoded particles having small color dots may beformed by sequential UV exposure under various magnetic fields. In thiscase, possible expressions of graphical codes are not limited due to theflexibility of controlling of colors and shapes.

According to one exemplary embodiment, a color encoded magneticstructure including a solid medium and a code region is provided. Thesolid medium may be a cured polymer. The code region may be present inthe solid medium. The code region may include magnetic nanoparticlesaligned in a chain structure. The magnetic nanoparticles may include asuperparamagnetic material. By the method described above with respectto the above exemplary embodiments, structural colors of the code regionmay be determined by inter-particle distance between the alignedmagnetic nanoparticles.

When the magnetic nanoparticles are aligned in various ways, chainstructures with various structural colors may be formed. Therefore, itis possible to implement multilevel coding using only one kind ofmagnetic nanoparticles. For example, multilevel codes more than binarycodes composed only of black and white colors may be easily realized.For example, quaternary, octal or hexadecimal codes composed of 4, 8 or16 colors may be realized.

The code region may include various color code information such asshapes or positions of color bits besides colors. For example, theshapes can be dots, lines or any arbitrary shapes.

The magnetic structure, for example, may be in the form of amicroparticle having a size of several tens to hundreds of micrometers.FIG. 5 shows images of various types of color encoded microparticles.Scale bars are 200 μm for A and B, 500 μm for C and 250 μm for D and E,respectively. As shown in A, C and D of FIG. 5, microparticles withvarious shapes and colors may be formed. In FIG. 5, B and E aretransmission electron microscope (TEM) images of samples correspondingto A and D, respectively. Unlike A and D having colorful reflectiveimages, B and E have brownish transmission images, which indicates thatthe color of the microparticles is caused by a structure of thesuperparamagnetic nanomaterial, not by coloring.

According to the color encoding by the above-mentioned method, severalcolors are disposed in local spaces separate from each other,respectively. In addition, a spectrum of the structural color in thelocal space has a single peak value. Thus, for example, position andcolor data of each code can be simultaneously obtained from positiondata and RGB data of a pixel only using a cheap charge coupled device(CCD) camera.

FIG. 6 shows images comparing a conventional binary encoded particlewith a color encoded particle. Binary encoding was performed byconcavo-convex patterning in 10 regions in a particle. Color encoding ina particle was performed by using 8 easily distinguishable colors toimplement multilevel coding. Referring to FIG. 6, in the left imageshowing binary encoding, only 2¹⁰ (≈10³) codes can be created with 10distinct bits. However, in the right image showing color encoding, 8¹⁰(=2³⁰≈10⁹) easily readable color codes can be created using a 10 bitsystem. This huge coding capacity of the color encoded particles enablesthe encoding of much larger molecular libraries with smaller particlesizes.

For multiplexed biomolecule assay, the color encoded magnetic structuremay further include a probe region. The probe region may include abinding site capable of binding an external target. The external targetis not particularly limited, but may include DNA, protein, or abiochemical material.

According to an exemplary embodiment, a method of analyzing a colorcode, which includes: providing a magnetic structure including a proberegion and a code region encoded with a color code; binding a targetwith the probe region; and decoding information from the magneticstructure with the target, wherein the code region includes a photoniccrystal of magnetic nanoparticles. The photonic crystal includes alignedmagnetic nanoparticles, and structural colors may be determined byinter-particle distance between the aligned magnetic nanoparticles. Inaddition, the information may include a shape, a position and a color ofthe color code, independently or in combination. The code region mayinclude at least one color code information selected from the groupconsisting of a shape, a position and a color of the color code.

FIGS. 7 to 10 show examples of a multiplexed biomolecule assay using acolor encoded magnetic structure to detect and identify DNAhybridization. All scale bars are 100 μm. The color encoded magneticstructure may have a smaller particle size than a typical probe spot ofa DNA microarray, and thus an encoding capacity may be more than severalbillions of kinds of possibilities.

FIG. 7 is a microscope image of a color encoded magnetic structure. Asshown in FIG. 7, a color encoded particle may include a code region andan oligonucleotide probe region. To avoid spectral overlapping of afluorescent signal for detection of hybridization with a structuralcolor signal for color encoding, the oligonucleotide probe region wasspatially separated from the code region. The code region wassynthesized from a composition for forming a color encoded magneticstructure, and the probe region was formed from a mixture of PEG-DA anda buffer solution of an acrylate-modified DNA oligomer probe.

FIG. 8 is reflective microscope images of multiplexed assays based onhybridization of three kinds of encoded particles and DNA oligomertargets. As shown in FIG. 8, 12.5 μM of DNA probes with differentnucleotides sequences (probe 1: 5′-ACA CTC TAC AAC TTC-3′, probe 2:5′-ATC AGA TTG GTT AGT-3′ and Control: a sample without a DNA probe)were incorporated into different color coded microparticles. 1 μM of DNAoligomer targets labeled with a fluorescent material were thenintroduced and incubated for 10 minutes. T1 and T2 are targetscomplementary to probe 1 and probe 2, respectively, a mark (+) indicatesthe presence of the target, and a mark (−) indicates the absence of thetarget. As a result, only the particles with DNA probes complementary tothe DNA oligomer targets show fluorescence.

The decoding process is simple and compatible with ordinary microscopesand color image analyzers. Decoding of a single encoded particle fortarget molecule identification is shown in FIGS. 9 and 10. FIG. 9 showsa result obtained after an encoded particle having probe 1 is hybridizedwith a complementary oligomer target (T1). FIG. 10 shows a resultobtained after the encoded particle of FIG. 9 is decoded. RGB levels ofindividual code positions were obtained from reflective micrographreadings using a full color CCD. The resulting RGB values were digitizedto specific code levels. Referring to FIG. 10, the respective RGB valueswere divided into 4 levels, resulting in four possibilities for each R,G and B value. The capability for multiplexing is virtually limitless.Here, for example, encoding and decoding of a microparticle withsquare-shaped color bits with 10 code positions and 4 color variationswas chosed.

According to the method of manufacturing a color encoded magneticstructure described above, the encoding capacity can be increased intothe billions using brilliant, easily distinguishable color encoding. Inaddition, the use of color tunable magnetic structure also enablescost-effective and scalable manufacturing of the color encodedmicroparticles by eliminating need of multiple coloring materials.Moreover, colors are magnetically tunable and lithographically fixable,and therefore high-definition patterning can be performed. Variousapplications can be developed by the above method. The above-mentionedcolor encoded magnetic structure can be useful in reflective displays,practical multiple bioassay and anti-counterfeiting of small objects.

The foregoing is illustrative of the present disclosure and is not to beconstrued as limiting thereof. Although numerous embodiments of thepresent disclosure have been described, those skilled in the art willreadily appreciate that many modifications are possible in theembodiments without materially departing from the novel teachings andadvantages of the present disclosure. Accordingly, all suchmodifications are intended to be included within the scope of thepresent disclosure as defined in the claims. Therefore, it is to beunderstood that the foregoing is illustrative of the present disclosureand is not to be construed as limited to the specific embodimentsdisclosed, and that modifications to the disclosed embodiments, as wellas other embodiments, are intended to be included within the scope ofthe appended claims. The present disclosure is defined by the followingclaims, with equivalents of the claims to be included therein.

What is claimed is:
 1. A method of analyzing a color code, comprising:providing a magnetic structure including a probe region and a coderegion encoded with a plurality of color codes which are fixed and havevarious colors the magnetic structure generated by sequentiallyirradiating patterned ultraviolet rays while i) varying a magnetic fieldstrength applied to the code region and ii) changing mask patterns sothat different regions of the code region are sequentially solidifiedwith different colors and different shapes; binding a target with theprobe region; and decoding information from the magnetic structure withthe target, wherein the code region includes a photonic crystal of fixedmagnetic particles.
 2. The method according to claim 1, wherein thefixed magnetic nanoparticles are aligned, and structural colors of thecode region are determined by inter-particle distance between the fixedmagnetic nanoparticles which are aligned.
 3. The method according toclaim 1, wherein the information includes at least one selected from thegroup consisting of a shape, a position and a color of the color code.4. The method of claim 1, wherein the code region is synthesized from acomposition including a curable material in which a plurality ofmagnetic nanoparticles are dispersed, wherein the composition issolidified so that the plurality of magnetic nanoparticles aredispersed, wherein the composition is solidified so that the pluralityof magnetic nanoparticles are fixed in the code region.
 5. The methodaccording to claim 1, wherein the decoding is performed by using amicroscope and an image analyzer.
 6. The method according to claim 5,wherein the decoding comprises obtaining RGB values of the individualcolor codes from micrograph readings.
 7. The method according to claim6, wherein the decoding further comprises digitizing the RGB values tospecific code levels.